Publications
| Title | Abstract | Year Filter | PMID(sorted ascending) Filter |
|---|
| the purification and properties of urocanase from pseudomonas testosteroni. | urocanase (urocanate hydratase, ec 4.2.1.49) purified from pseudomonas testosteroni has a mol.wt. of 118000 determined by sedimentation-equilibrium analysis. ultracentrifugation in 6m-guanidine hydrochloride and polyacrylamide-gel electrophoresis in sodium dodecyl sulphate show that the enzyme consists of two identical or very similar subunits. it is, like urocanase isolated from other sources, inhibited by reagents that react with carbonyl groups. although urocanase from ps. testosteroni is str ... | 1978 | 25660 |
| phthalate and 4-hydroxyphthalate metabolism in pseudomonas testosteroni: purification and properties of 4,5-dihydroxyphthalate decarboxylase. | phthalate is degraded through 4,5-dihydroxyphthalate and protocatechuate in pseudomonas testosteroni nh1000. the ezyme 4,5-dihydroxyphthalate decarboxylase, catalyzing the conversion of 4,5-dihydroxyphthalate to protocatechuate and carbon dioxide, was purified approximately 130-fold from phthalate-induced cells of a protocatechuate 4,5-dioxygenase-deficient mutant of p. testosteroni. the most purified preparation showed a single protein band on sodium dodecyl sulfate-acrylamide disc gel electrop ... | 1978 | 29563 |
| binding of steroids by a partially purified periplasmic protein from pseudomonas testosteroni. | 1979 | 41976 | |
| effect of testosterone-estradiol-binding globulin in the enzymic oxidoreduction of 17-oxygenated c19 steroids. | the effect of both testosterone-estradiol-binding globulin (tebg) and albumin on enzymic oxidoreduction of four 17-oxygenated c19 steroids by bacterial 17beta-hydroxysteroid:nad oxidoreductase from pseudomonas testosteroni was investigated. the decreased yields of products under presence of tebg were found in both directions of reversible enzymic reaction. this finding was unexpected in the case of enzymic reduction in which the opposite effect could be assumed with respect of high affinity of t ... | 1975 | 50994 |
| affinity partition of proteins in aqueous two-phase systems containing polyoxyethylene glycol-bound ligand and charged dextrans. | the partition of the delta 5 leads to 4 3-oxosteroid isomerase of pseudomonas testosteroni in aqueous two-phase systems containing both the macroligand, polyoxyethylene glycol-bound estradiol, and charged (cationic or anionic) dextrans has been studied. if the enzyme is well retained in the upper phase by an adequate amount of macroligand, it is possible to improve the removal of the contaminating proteins by extracting them into a lower phase containing positively or negatively charged dextran. ... | 1979 | 94912 |
| [chromosomal structures of pseudomonas testosteroni. i. isolation and characterization of the chromosomal complexes. (author's transl)]. | after lysis of pseydomonas testosteroni with lysozyme and non-ionic detergents different dna-protein complexes can be separated in 5-25% (w/v) neutral sucrose gradient. the protein to dna ratio of these complexes varies between 0.5-4.5 to 1, whereby the faster sedimenting forms contain more protein than the slower sedimenting ones. different initial rates of dnase digestion may indicate various degrees of dna packing in these complexes. the chromosomal complexes of pseudomonas testosteroni are r ... | 1976 | 132046 |
| [chromosomal structure of pseudomonas testosteroni. ii. activity of the endogenous rna-polymerase (author's transl)]. | after careful lysis the nucleoid of pseudomonas testosteroni can be isolated in three different forms with compact and unfolded dna structures. the released nucleoids contain endogenous dna-dependent rna-polymerase activity using the chromosomal dna as a template. rna synthesis is proportional to duration of rna-polymerase reaction and amount of dna-protein-complexes. the sensitivity towards ionic strength and rifampicin indicates that a part of rna-polymerase activity is tightly bound to the ch ... | 1976 | 136130 |
| [chromosomal structures of pseudomonas testosteroni. iv. effect of testosterone on rna-synthesis (author's transl)]. | testosterone degrading enzymes are synthesized de novo by bacterium p. testosteroni to utilize testosterone-like steroids as the only source of carbon. rna-synthesis of the whole lysate of testosterone-induced bacteria was found to be 15% reduced compared to the control, suggesting a cytoplasmatic factor which modulates chromatin associated rna-polymerase activity. | 1977 | 143827 |
| bacterial metabolism of arylsulfonates: role of meta cleavage in benzene sulfonate oxidation by pseudomonas testosteroni. | pseudomonas testosteroni h-8 oxidizes certain lower alkylbenzene sulfonates at rates inversely related to the length of the alkyl group. appreciable q(o)2 values were observed for benzene sulfonate (bs), toluene sulfonate (ts), and ethylbenzene sulfonate (ebs), but not for propylbenzene sulfonate (ps) and higher homologues. catechol oxidation was catalyzed by a constitutive catechol-2,3-oxygenase (ec 1.99.2.a). yellow meta cleavage products accumulated when bs-grown cells were exposed to catecho ... | 1975 | 163618 |
| enzymatic transformation of morphine by hydroxysteroid dehydrogenase from pseudomonas testosteroni. | eznyme preparations from pseudomonas testosteroni containing alpha- and beta- hydroxysteroid dehydrogenases catalyzed the oxidation of morphine and codeine by nicotinamide adenine dinucleotide. morphine was converted in relatively low yield into 14-hydroxymorphinone probably via morphinone as an intermediate. codeine was converted to codeinone and 14-hydroxycodeinone. only the conversions at the 6-position were carred out by the hydroxysteroid dehydrogenase. hydroxylation at the 14-position did ... | 1975 | 172013 |
| horse-liver alcohol dehydrogenase and pseudomonas testosteroni 3(17)beta-hydroxysteroid dehydrogenase transfer epimeric hydrogens from nadh to 17beta-hydroxy-5alpha-androstan-3-one. an exception to one of the alworth-bentley rules. | in the reduction of 17beta-hydroxy-5alpha-androstan-3-one to the 3beta-alcohol, horse liver alcohol dehydrogenase utilizes the 4-pro-r hydrogen of nadh whereas the 3(17)beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni utulized the 4-pro-s hydrogen. these observations provide an exception to the rule proposed by alworth and bentley that with regard to the paired methylene hydrogens at c-4 of nadh and nadph "the stereospecificity of a particular reaction is fixed and does not vary wit ... | 1976 | 177288 |
| membrane bound 3beta and 17beta-hydroxysteroid dehydrogenase and its role in steroid transport in membrane vesicles of pseudomonas testosteroni. | 1976 | 180333 | |
| affinity chromatography of 3 alpha-hydroxysteroid dehydrogenase from pseudomonas testosteroni. use of n,n-dimethylformamide to prevent hydrophobic interactions between the enzyme and the ligand. | 1. the 3alpha-hydroxysteroid: nad+-oxidoreductase (ec 1.1.1.50) from pseudomonas testosteroni (atcc 11996) has been purified by affinity chromatography on sepharose 4b using glycocholic acid as ligand covalently bound through its carboxyl group to the ethylenediamine spacer. 2. the attachment of the enzyme to the substrate-containing matrix is greatly enhanced by the presence of nad+ suggesting that this enzyme has a compulsory ordered mechanism where nad+ binds to the enzyme before the steroid. ... | 1976 | 181083 |
| 3(17)beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni. a convenient purification and demonstration of multiple molecular forms. | 1977 | 193845 | |
| 3(17)beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni. ligand binding properties. | the binding of nad and nadh to electrophoretically pure 3(17)beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni was determined by fluorescence spectroscopy and gel filtration. four moles of cofactor are bound/mol of tetrameric enzyme; the binding sites are equivalent and independent. the dissociation constants for nad and nadh are 16 and 0.25 micronm, respectively. as measured by gel filtration in the absence of cofactor, 0.4 mol of estradiol-17 beta is bound/mol of tetrameric enzyme. ... | 1977 | 193846 |
| 3alpha-hydroxysteroid dehydrogenase from pseudomonas testosteroni: kinetic properties with nad and its thionicotinamide analogue. | the kinetics of 3alpha-hydroxysteroid : nad oxidoreductase (ec 1.1.1.50) from pseudomonas testosteroni (atcc 11996) have been investigated. the kinetic analysis based on initial activity measurements and product inhibition studies, indicates that the addition of substrate to the enzyme and the release of products from it, follows an obligatory order (ordered bi bi mechanism). the ability of the enzyme to utilize the thionicotinamide analogue of nad (snad) as cofactor has been investigated using ... | 1975 | 234846 |
| allantoin racemase: a new enzyme from pseudomonas species. | 1. allantoin racemase is a novel enzyme which catalyzes the conversion of s(+)-and r(minus)-allantoin into the racemate. 2. the enzyme is present in pseudomonas testosteroni, pseudomonas putida and five biotypes of pseudomonas fluorescens, but absent in a number of other pseudomonas species. 3. the enzyme of ps. testosteroni was purified 133-fold and exposes optimal activity at ph 8.0-8.2 and 50 degrees c. the enzyme is stable on heating for 15 min at 70 degrees c. 4. the enzyme appeared to be s ... | 1975 | 237557 |
| effect of protein concentration on the molecular weight of delta5-3-ketosteroid isomerase. | the molecular weight of delta-5-3-ketosteroid isomerase from pseudomonas testosteroni was determined by means of sedimentation equilibrium and exclusion chromatography over a wide range of enzyme concentrations in 0.2 m potassium phosphate buffer, ph 7.0. in addition, the sedimentation constant of the enzyme was determinded over an extended range of concentrations. the enzyme was found to have a molecular weight of 26,000 plus or equal to 1,000, suggesting that it is a dimer of identical or simi ... | 1975 | 237889 |
| purification and properties of malate dehydrogenase from pseudomonas testosteroni. | nicotinamide adenine dinucleotide-linked malate dehydrogenase has been purified from pseudomonas testosteroni (atcc 11996). the purification represents over 450-fold increase in specific activity. the amino acid composition of the enzyme was determined and found to be quite different from the composition of the malate dehydrogenases from animal sources as well as from escherichia coli. despite this difference, however, the data show that the enzymatic properties of the purified enzyme are remark ... | 1975 | 238957 |
| the purification and properties of l-histidine--2-oxoglutarate aminotransferase from pseudomonas testosteroni. | 1. inducible l-histidine--2-oxoglutarate aminotransferase was purified some 170-fold from extracts of pseudomonas testosteroni. 2. the preparation showed only one major component after electrophoresis on polyacrylamide gels, though additional minor bands were observed when samples concentrated on a deae-cellulose column were used. 3. the molecular weight of the enzyme was found to be approx. 70000 by chromatography on sephadex g-200. 4. the purification scheme produced enzyme that was inactive i ... | 1975 | 241324 |
| specific gonadotropin binding to pseudomonas maltophilia. | binding of 125i-labeled human chorionic gonadotropin to pseudomonas maltophilia is dependent on time, temperature, and ph and the binding to this procaryotic species is hormone-specific and saturable. the equilibrium dissociation constant is 2.3 x 10(-9) m. there are no cooperative interactions between binding sites (hill coefficient, 1.05). the number of sites is estimaated as 240 fmol/100 mug of protein. nacl and kcl, at concentrations from 1 to 10 mm, have no effect on binding. divalent catio ... | 1977 | 265583 |
| purine degradation in pseudomonas aeruginosa and pseudomonas testosteroni. | 1. adenine, hypoxanthine, xanthine and guanine are broken down in pseudomonas aeruginosa and pseudomonas testosteroni to allantoin by the concerted action of the enzymes adenine deaminase, guanine deaminase, nad+-dependent xanthine dehydrogenase and uricase. 2. uric acid is broken down by an unstable, membrane-bound uricase with an unusually low ph optimum. 3. in both strains adenine inhibits growth and xanthine dehydrogenase. a second type of inhibition is manifest only in ps. testosteroni and ... | 1977 | 407941 |
| hydroxysteroid dehydrogenases of pseudomonas testosteroni. separation of a 17 beta-hydroxysteroid dehydrogenase from the 3(17) beta-hydroxysteroid dehydrogenase and comparison of the two enzymes. | when a crude extract of pseudomonas testosteroni induced with testosterone was subjected to polyacrylamide gel electrophoresis, six bands that stained for 17 beta-hydroxysteroid dehydrogenase activity was observed. a protein fraction containing the enzyme corresponding to the fastest migrating band and devoid of the other hydroxysteroid dehydrogenase activities has been obtained. this preparation appears to be distinct from the previously isolated 3(17) beta-hydroxysteroid dehydrogenase (ec 1.1. ... | 1977 | 411517 |
| "affinity" chromatography of steroid-transforming enzymes with a non-steroidal ligand. | the chromatographic behaviour of an avian oestradiol-17 beta dehydrogenase, the 3(17) beta-hydroxy steroid dehydrogenase from pseudomonas testosteroni and cortisone reductase from streptomyces dehydrogenans was studied on columns of p-(phenoxypropoxy)aniline attached to cnbr-activated sepharose. the ligand was effective in adsorbing the oestradiol dehydrogenase from a partially purified extract of chicken liver, and the cortisone reductase was perferentially retained when mixtures of the three d ... | 1979 | 435242 |
| phosphate and soil binding: factors limiting bacterial degradation of ionic phosphorus-containing pesticide metabolites. | soils that had a high binding capacity for inorganic orthophosphate (pi) had reduced capacities to bind ionic alkyl phosphorus compounds. only ionic methylphosphonate (mpn) and ionic phenylphosphonate exhibited moderate binding. pseudomonas testosteroni used either mpn or pi as a sole phosphorus source and exhibited diauxic utilization of mpn and pi. the utilization of mpn was suppressed in the presence of pi. this suppression was abolished by a pi-binding soil. the soil did not have a significa ... | 1979 | 453832 |
| the effects of specific inhibitors and an antiserum of 3 beta and 17 beta-hydroxysteroid dehydrogenase of steroid uptake in pseudomonas testosteroni. | 1979 | 459502 | |
| localization of 3 beta and 17 beta-hydroxysteroid dehydrogenase in pseudomonas testosteroni. | 1979 | 459503 | |
| inhibition of the 3 beta-hydroxysteroid oxidoreductase of pseudomonas testosteroni by steroids. | 55 steroids of the estratriene and androstane type with substituents in pos. 16 alpha, 17 alpha or 17 beta were tested for inhibition of the 3beta-hydroxysteroid oxidoreductase of pseudomonas testosteroni. estratrien-3-ols were strong and competitive inhibitors (ki less than 1 micron). substituents in pos. 16 alpha of estradiol influenced the inhibitory activity distinctly. substituents in 17 alpha- or 17 beta-position were of slight influence. 3-methoxy estratrienes gave no inhibition of the en ... | 1979 | 467367 |
| the amino acid composition of histidine ammonialyase from pseudomonas putida ncib 10807. | the amino acid composition of histidine ammonia-lyase from pseudomonas putida ncib 10807 suggests that this enzyme may be different from the pseudomonas testosteroni ncib 10808 histidine ammonia-lyase, whose amino acid composition is known. | 1979 | 488259 |
| characterisation of an associate 17-beta-hydroxysteroid dehydrogenase activity and affinity labelling of the 3-alpha-hydroxysteroid dehydrogenase of pseudomonas testosteroni. | the 3-alpha-hydroxysteroid dehydrogenase and the 3-beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni were purified to homogeneity by polyaerylamide gel electrophoresis using the following stages: deae cellulose chromatography, affinity chromatography on oestrone-aminocaproate sepharose and sephadex gel filtration. the pure 3-alpha-hydroxysteroid dehydrogenase was completely devoid of 3-beta-hydroxysteroid dehydrogenase activity but could oxidize estradiol 17-beta at an appreciable ra ... | 1977 | 607995 |
| continuous dehydrogenation of a steroid with immobilized microbial cells: effect of an exogenous electron acceptor. | whole cells of pseudomonas testosteroni, induced to synthesize steroid-transforming enzymes beforehand, have been immobilized by entrapment in polyacrylamide gel. the immobilized cells have been used to catalyze the continuous delta1-dehydrogenation of reichstein's substance s under various conditions in the presence of phenazine methosulfate (pms), an electron acceptor for the cell-free delta1-dehydrogenase. the presence of pms substantially increases the rate of reaction when fed with the ster ... | 1978 | 623902 |
| formation of 5-[17beta-2h]androstene-3beta,17alpha-diol from 3beta-hydroxy-5-[17,21,21,21-2h]pregnen-20-one by the microsomal fraction of boar testis. | after incubation of 3beta-hydroxy-5-[17,21,21,21-2h]-pregnen-20-one with the microsomal fraction of boar testis, the metabolites were analyzed by gas chromatography and gas chromatography-mass spectrometry. the following metabolites were identified: 3beta,17alpha-dihydroxy-5-[21,21,21-3h]pregnen-20-one, 3beta-hydroxy-5-androsten-17-one, 5-androstene-3beta,17beta-diol, and 5-[17beta-2h]androstene-3beta,17alpha-diol. the presence of a 2h atom at the 17beta position of 5-androstene-3beta,17alpha-di ... | 1978 | 659416 |
| phthalate metabolism in pseudomonas testosteroni: accumulation of 4,5-dihydroxyphthalate by a mutant strain. | a mutant strain of pseudomonas testosteroni blocked in phthalate catabolism converted phthalate into 4,5-dihydroxyphthalate. the latter compound was isolated, and its physical properties were determined. a stoichiometric conversion of the compound to protocatechuate was demonstrated spectrophotometrically with crude extracts of a protocatechuate 4,5-dioxygenase-deficient mutant. therefore, phthalate is metabolized through 4,5-dihydroxyphthalate and protocatechuate, which is further degraded by p ... | 1977 | 873893 |
| interaction of steroids with pseudomonas testosteroni 3-oxosteroid delta4--delta5-isomerase. | 1977 | 911812 | |
| a reinvestigation of the mechanism of pseudomonas testosteroni delta 5-3-ketosteroid isomerase. | the mechanism of the isomerisation of delta 5-3,17-androstenedione by the isomerase (3-oxosteroid delta 4-delta 5-isomerase, ec 5.3.3.1) of pseudomonas testosteroni has been reinvestigated with delta 5-[4-beta-2h]androstenedione as substrate in h2o and delta 5-androstenedione in 2h2o. a precise localisation of the label in delta 4-androstenendione has revealed that the previously reported 4 beta leads to 6 beta deuterium transfer accounts for only a part of the reaction. along with this process ... | 1977 | 922021 |
| effect of sulfhydryl and disulfide agents on 3beta and 17beta-hydroxysteroid dehydrogenase and on steroid uptake of pseudomonas testosteroni. | 1976 | 966766 | |
| temperature-sensitive rna synthesis during adaptive growth on testosterone of pseudomonas testosteroni. | 1976 | 979269 | |
| the involvement of the electron transport chain in uptake of testosterone by membrane vesicles of pseudomonas testosteroni. | 1976 | 1011843 | |
| concentration-dependent association of delta5-3-ketosteroid isomerase of pseudomonas testosteroni. | gel chromatography and ultracentrifugation studies show that delta5-3-ketosteroid isomerase of pseudomonas testosteroni a dimer with a molecular weight of 26,800 at concentrations below 1 mg per ml, undergoes reversible, concentration-dependent association at higher enzyme concentrations. in the concentration range between 0.04 and 15.6 mg per ml, apparent molecular radii of 23 a to 36 a and molecular weights of 26,000 to 69,000 were observed. the latter value represents the weight average molec ... | 1975 | 1141206 |
| specificity of oxidation of bile-salt hydroxyl groups by crude extracts of pseudomonas testosteroni (atcc 11996) used in determining bile salts. | recent experimental evidence suggests that, presumably as a recent of mutation, crude extracts of pseudomonas testosteroni (atcc 11996) no contain amounts of 7alpha- and 12alpha-hydroxysteroid dehydrogenate activity that invalidate the results of bile-salt determinations. to confirm or deny this, we studied the specificity of hydroxysteroid dehydrogenase contained in crude extracts of currently available samples of this bacterium in the oxidation of bile-salt hydroxyl groups. the dehydrogenases ... | 1975 | 1164792 |
| chemical modification of amino acid residues associated with the delta-4-3-ketosteroid-dependent photoinactivation of delta-5-3-ketosteroid isomerase. | we have studied the accumulation of dibenzyldimethyl-ammonium ion (dda+) by respiring membrane vesicles of escherichia coli, as an index of the generation of an electrical gradient during respiration. nonrespiring vesicles accumulated dda+ when k+ efflux was induced by valinomycin or monactin. by various criteria this was shown to be the exchange of one cation for another, independent of metabolism and coupled entirely by electrical forces. uptake of dda+ by respiring vesicles was inhibited by i ... | 1975 | 1167545 |
| letter: pseudomonas testosteroni septicemia. | 1975 | 1180436 | |
| on the 3alpha-hydroxysteroid dehydrogenase from pseudomonas testosteroni. the effect of denaturing agents. | highly purified preparations of the 3alpha-hydroxysteroid:nad- oxidoreductase (e.c.1.1.1.50) from pseudomonas testosteroni (atcc 11996) which consist of two major isoenzymes, with traces of a third, have been split into two enzymatically inactive polypeptides a and b by the use of sodium dodecylsulphate, urea and guanidinium hydrochloride. both polypeptides have a molecular weight of 25,000 +/- 2,500 as shown by thin-layer gel chromatography and ultracentrifugations. they differ, however, in cha ... | 1975 | 1184284 |
| substrate polarization by residues in delta 5-3-ketosteroid isomerase probed by site-directed mutagenesis and uv resonance raman spectroscopy. | delta 5-3-ketosteroid isomerase (ksi: ec 5.3.3.1) of pseudomonas testosteroni catalyzes the isomerization of delta 5-3-ketosteroids to delta 4-3-ketosteroids by the stereospecific transfer of the steroid 4 beta-proton to the 6 beta-position, using tyr-14 as a general acid and asp-38 as a base. ultraviolet resonance raman (uvrr) spectra have been obtained for the catalytically active double mutant y55f + y88f, which retains tyr-14 as the only tyrosine residue (referred to as the y14(0) mutant), a ... | 1992 | 1339027 |
| broad host-range vector for efficient expression of foreign genes in gram-negative bacteria. | a broad host-range expression plasmid was constructed comprising the incq replicon, the reca promoter from escherichia coli and the g10-l ribosome binding site (rbs) derived from bacteriophage t7. the structural genes for porcine somatotropin (pst) and e. coli beta-galactosidase (lacz) were used to monitor gene expression in a diverse collection of gram-negative bacterial hosts: escherichia coli, pseudomonas aeruginosa, pseudomonas syringae, pseudomonas putida, pseudomonas fluorescens, pseudomon ... | 1991 | 1367537 |
| a novel sulfatase from pseudomonas testosteroni hydrolyzing lithocholic acid sulfate. | pseudomonas testosteroni atcc 11996 was found to produce a novel bile acid sulfate sulfatase that hydrolyzes the sulfate ester bond in lithocholic acid sulfate (lca-s). the enzyme synthesis was induced by several kinds of bile acids including lca-s. mn2+ functioned as an essential component for the enzyme synthesis and so4(2-) suppressed it. this sulfatase hydrolyzes lca-s to isolithocholic acid and sulfuric acid with inversion of alpha- to beta-configuration of the hydroxyl group at the third p ... | 1992 | 1369058 |
| change of speciation of cu(ii) in growth medium due to uptake of ammonia by pseudomonas testosteroni during growth. | growth of pseudomonas testosteroni in a medium containing 1 mm cu(ii) causes a color change from blue to green. the spectrum of the supernatant solution from the blue culture shows an absorption at 660 nm, identical to that of 1 mm [cu(ii)] in the medium. the green supernatant solution shows a uv absorption, which tails into the visible and so is responsible for the green color, and a d-d absorption at 720 nm. the absorption at 660 nm for the blue supernatant solution is probably due to [cu(nh3) ... | 1992 | 1369191 |
| identification of xenobiotic-degrading isolates from the beta subclass of the proteobacteria by a polyphasic approach including 16s rrna partial sequencing. | nineteen gram-negative, aerobic, biodegradative isolates were identified by using a polyphasic taxonomic approach. the presence of the specific polyamine 2-hydroxyputrescine and the presence of a ubiquinone with eight isoprenoid units in the side chain (ubiquinone q-8) allowed allocation of these organisms to the beta subclass of the proteobacteria. on the basis of the results of additional characterization experiments (i.e., api 20ne tests, determinations of soluble protein patterns, and dna-dn ... | 1992 | 1371062 |
| kinetics and stability of delta 5-3-ketosteroid isomerase from pseudomonas testosteroni in the system of reverse micelles of aerosol ot in isooctane. | partially purified delta 5-3-ketosteroid isomerase (ksi) from pseudomonas testosteroni was studied kinetically after solubilization in reverse micelles of aerosol ot (aot) in isooctane and water, as regards its application to biotechnology. with delta 5,10-estren-17 beta-ol-3-one as a substrate, ksi displays an enzyme activity in the micellar system but a low stability. in the presence of urea, the enzyme is, however, stable. kinetic parameters of the stabilized enzyme are highly sensitive to bo ... | 1992 | 1381618 |
| homologies between enzymes involved in steroid and xenobiotic carbonyl reduction in vertebrates, invertebrates and procaryonts. | evidence is reported for the existence of a structurally and functionally related and probably evolutionarily conserved class of membrane-bound liver carbonyl reductases/hydroxysteroid dehydrogenases involved in steroid and xenobiotic carbonyl metabolism. carbonyl reduction was investigated in liver microsomes of 8 vertebrate species, as well as in insect larvae total homogenate and in purified 3 alpha-hydroxysteroid dehydrogenase preparations of the procaryont pseudomonas testosteroni, using th ... | 1992 | 1472459 |
| functional and immunological relationships between metyrapone reductase from mouse liver microsomes and 3 alpha-hydroxysteroid dehydrogenase from pseudomonas testosteroni. | 3 alpha-hydroxysteroid dehydrogenase (3 alpha-hsd) from pseudomonas testosteroni was shown to reduce the xenobiotic carbonyl compound metyrapone (mpon). reversely, mpon reductase purified from mouse liver microsomes and previously characterized as aldehyde reductase, was competitively inhibited by 3 alpha-hsd steroid substrates. for mpon reduction both enzymes can use either nadh or nadph as co-substrate. immunoblot analysis after native and sds gel electrophoresis of 3 alpha-hsd gave a specific ... | 1992 | 1551429 |
| specific activation of a tyrosine----glycine mutant of delta 5-3-ketosteroid isomerase by phenols. | a key unknown still to be explored concerning the mechanism of delta 5-3-ketosteroid isomerase from pseudomonas testosteroni is the extent of the proton transfer between tyrosine-14 of the enzyme and the c-3 carbonyl oxygen of the steroid substrate. this report is a preliminary study of a system we are developing to allow us eventually to use a brønsted analysis to measure this transfer. we describe the construction of an expression vector and tyrosine-14----glycine-14 mutant of the enzyme and i ... | 1992 | 1590799 |
| dye-linked dehydrogenase activities for formate and formate esters in amycolatopsis methanolica. characterization of a molybdoprotein enzyme active with formate esters and aldehydes. | cell-free extracts of methanol-grown amycolatopsis methanolica contain dye-linked dehydrogenase activities for formate and methyl formate. fractionation of the extracts revealed that the (unstable) activity for formate resides in membrane particles, while that for methyl formate belongs to a soluble enzyme that was purified and characterized. the enzyme, indicated as formate-ester dehydrogenase, appeared to be a molybdoprotein (4 fe, 3 or 4 s, 1 mo and 1 fad were found for each enzyme molecule), ... | 1992 | 1597191 |
| effects of chlorobenzoate transformation on the pseudomonas testosteroni biphenyl and chlorobiphenyl degradation pathway. | bacterial conversion of biphenyl (bp) and chlorobiphenyls (cbps) to benzoates and chlorobenzoates (cbas) proceeds by introduction of molecular oxygen at the 2,3 position, followed by a 1,2-meta cleavage of the molecule. complete mineralization of cbps requires the presence of two sets of genes, one for the transformation fo cbps into cbas and a second for the degradation of cbas. it has been shown previously that removal of the cbas produced from the degradation of cbps is essential for efficien ... | 1992 | 1610172 |
| modelling of mixed chemostat cultures of an aerobic bacterium, comamonas testosteroni, and an anaerobic bacterium, veillonella alcalescens: comparison with experimental data. | a mathematical model of mixed chemostat cultures of the obligately aerobic bacterium comamonas testosteroni and the anaerobic bacterium veillonella alcalescens grown under dual limitation of l-lactate and oxygen was constructed. the model was based on michaelis-menten-type kinetics for the consumption of substrates, with noncompetitive inhibition of v. alcalescens by o2. the growth characteristics of the aerobic and anaerobic organisms were determined experimentally with pure cultures of the ind ... | 1992 | 1622213 |
| outer membranes of gram-negative bacteria are permeable to steroid probes. | the permeability of bacterial outer membranes was assayed by coupling the influx of highly hydrophobic probes, 3-oxosteroids, with their subsequent oxidation catalysed by 3-oxosteroid delta 1-dehydrogenase, expressed from a gene cloned from pseudomonas testosteroni. in salmonella typhimurium producing wild-type lipopolysaccharide, the permeability coefficients for uncharged steroids were 0.45 to 1 x 10(-5) cm s-1, and the diffusion appeared to occur mainly through the lipid bilayer domains of th ... | 1992 | 1640833 |
| molecular cloning and expression of the beta-hydroxysteroid dehydrogenase gene from pseudomonas testosteroni. | the structural gene (hsd) of the pseudomonas testosteroni encoding the 17 beta-hydroxysteroid dehydrogenase has been cloned using the cosmid vector pvk102. escherichia coli carrying recombinant clones of hsd, isolated by immunological screening, were able to express the biologically active enzyme, as measured by the conversion of testosterone into androstenedione. subcloning experiments, restriction and deletion analysis, and site-directed insertion mutagenesis showed that the hsd gene is locate ... | 1991 | 1657714 |
| cloning, sequencing, and expression of the pseudomonas testosteroni gene encoding 3-oxosteroid delta 1-dehydrogenase. | pseudomonas testosteroni atcc 17410 is able to grow on testosterone. this strain was mutagenized by tn5, and 41 mutants defective in the utilization of testosterone were isolated. one of them, called mutant 06, expressed 3-oxosteroid delta 1- and 3-oxosteroid delta 4-5 alpha-dehydrogenases only at low levels. the dna region around the tn5 insertion in mutant 06 was cloned into puc19, and the 1-kbp ecori-bamhi segment neighbor to the tn5 insertion was used to probe dna from the wild-type strain. ... | 1991 | 1657885 |
| microbial degradation of the morphine alkaloids: identification of morphine as an intermediate in the metabolism of morphine by pseudomonas putida m10. | a strain of pseudomonas putida was isolated by selective enrichment with morphine that was capable of utilising morphine as a primary source of carbon and energy for growth. experiments with whole cells showed that both morphine and codeine, but not thebaine, could be utilised. a novel nadp-dependent dehydrogenase, morphine dehydrogenase, was purified from crude cell extracts and was shown to be capable of oxidising morphine and codeine to morphinone and codeinone, respectively. this nadp-depend ... | 1990 | 1701625 |
| carbonyl reduction of metyrapone in human liver. | carbonyl reduction was investigated in cytosolic and microsomal fractions of human liver using the ketone metyrapone as a substrate. the cytosolic enzyme has a stronger preference for nadph over nadh than the microsomal enzyme: the former shows only 14% of the nadph-supported activity while the latter exhibits 36% activity with nadh. barbitone and quercitrin, the classic inhibitors of carbonyl reductases, do not affect metyrapone reduction in either fraction. dicumarol and indomethacin, the spec ... | 1991 | 1722672 |
| subcloning of bph genes from pseudomonas testosteroni b-356 in pseudomonas putida and escherichia coli: evidence for dehalogenation during initial attack on chlorobiphenyls. | the bpha, -b, -c, and -d genes from pseudomonas testosteroni b-356 were mapped to a 5.5-kb dna fragment of cloned plasmids pda1 and pda2 by use of deletion and insertion mutants of these plasmids. the expression of each of these genes was evaluated in escherichia coli and in pseudomonas putida, and it was found that the bphc and bphd genes are well expressed in both e. coli and p. putida cells while the bpha and bphb genes are very poorly expressed in e. coli, even when placed downstream of a ta ... | 1991 | 1746948 |
| bioconversion of 2-hydroxy-6-oxo-6-(4'-chlorophenyl)hexa-2,4-dienoic acid, the meta-cleavage product of 4-chlorobiphenyl. | bacterial conversion of 4-chlorobiphenyl (4-cb) usually proceeds through a pathway involving an initial oxidation of the unsubstituted ring in the 2,3 position followed by a 1,2 meta-cleavage. the meta-cleavage product (mcp) is converted through a single hydrolysis step into chlorobenzoic acid. however, several other acidic metabolites that were not expected as part of this pathway have already been described. in this paper, we used strains of pseudomonas putida carrying cloned genes from pseudo ... | 1991 | 1919512 |
| studies of the catalytic mechanism of an active-site mutant (y14f) of delta 5-3-ketosteroid isomerase by kinetic deuterium isotope effects. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereoselective transfer of the 4 beta-proton to the 6 beta-position. the rate-limiting step has been shown to be the concerted enolization of the enzyme-bound substrate comprising protonation of the 3-carbonyl oxygen by tyr-14 and abstraction of the 4 beta-proton by asp-38 [xue, l., talalay, p., & mildvan, a. s. (1990) biochemistry 29, ... | 1991 | 1932008 |
| 4-sulphobenzoate 3,4-dioxygenase. purification and properties of a desulphonative two-component enzyme system from comamonas testosteroni t-2. | cell-free extracts of comamonas testosteroni t-2 grown in toluene-p-sulphonate/salts medium catalyse the conversion of p-sulphobenzoate (psb) into protocatechuate and sulphite by an nadh-requiring and fe2(+)-activated dioxygenase. anion-exchange chromatography of extracts yielded red (a) and yellow (b) protein fractions, both of which were necessary for dioxygenative activity. further purification of each fraction by hydrophobic interaction chromatography and gel filtration led to two homogeneou ... | 1991 | 2012609 |
| pseudomonas 3 beta-hydroxysteroid dehydrogenase. primary structure and relationships to other steroid dehydrogenases. | the 3 beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni commercially available was purified by an fplc step and submitted to sequence determination by peptide analysis. the structure obtained reveals a 253-residue polypeptide chain, with an n-terminal, free alpha-amino group, and a low cysteine content. comparisons with other hydroxysteroid dehydrogenases recently characterized reveal distant similarities with prokaryotic and, to some extent, also eukaryotic forms of separate specifi ... | 1991 | 2026158 |
| catalytic mechanism of an active-site mutant (d38n) of delta 5-3-ketosteroid isomerase. direct spectroscopic evidence for dienol intermediates. | the delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni catalyzes the conversion of androst-5-ene-3,17-dione to androst-4-ene-3,17-dione by a stereospecific transfer of the 4 beta-proton to the 6 beta-position. the reaction involves two steps: (a) a rate-limiting concerted enolization, comprising protonation of the 3-carbonyl oxygen by the phenolic hydroxyl group of tyr-14 and abstraction of the 4 beta-proton by the carboxylate group of asp-38, and (b) rapid reketonization o ... | 1991 | 2036366 |
| 4-toluene sulfonate methyl-monooxygenase from comamonas testosteroni t-2: purification and some properties of the oxygenase component. | comamonas testosteroni t-2 synthesizes an inducible enzyme system that oxygenates 4-toluene sulfontate (ts) to 4-sulfobenzyl alcohol when grown in ts-salts medium. we purified this ts methyl-monooxygenase system (tsmos) and found it to consist of two components. a monomeric, iron-sulfur flavoprotein (component b), which has been shown to act as a reductase in the 4-sulfobenzoate dioxygenase system of this organism (h. h. locher, t. leisinger, and a. m. cook, biochem. j. 274:833-842, 1991), carri ... | 1991 | 2050632 |
| beta-hydroxysteroid dehydrogenase: activity in microemulsion and extraction from pseudomonas testosteroni cells with microemulsion. | the stability of purified beta-hydroxysteroid dehydrogenase activity measured as a function of time was good in buffered cationic and non-ionic microemulsions. the use of 1-pentanol and 1-hexanol in place of 1-butanol as cosurfactant gave increased activity and stability. the nad+ michaelis constant was 0.22 mm in buffer and 3.5 mm in waterpool concentration in microemulsion. proteins, among them beta-hydroxysteroid dehydrogenase, were extracted from pseudomonas testosteroni with cationic microe ... | 1990 | 2116917 |
| chlorinated biphenyl mineralization by individual populations and consortia of freshwater bacteria. | comparative studies were performed to investigate the contribution of microbial consortia, individual microbial populations, and specific plasmids to chlorinated biphenyl biodegradation among microbial communities from a polychlorinated biphenyl-contaminated freshwater environment. a bacterial consortium, designated lps10, was shown to mineralize 4-chlorobiphenyl (4cb) and dehalogenate 4,4'-dichlorobiphenyl. the lps10 consortium involved three isolates: pseudomonas testosteroni (lps10a), which m ... | 1990 | 2117875 |
| quinoprotein alcohol dehydrogenase from pseudomonas aeruginosa and quinohemoprotein alcohol dehydrogenase from pseudomonas testosteroni. | 1990 | 2126332 | |
| induction of il-1 during hemodialysis: transmembrane passage of intact endotoxins (lps). | circulating monocytes of patients undergoing chronic hemodialysis are triggered to produce interleukin-1 (il-1) in vivo. intradialytic induction of il-1 is associated with complement activation in patients dialyzed with first-use cellulose membranes. chronic stimulation of il-1 production occurs because of an yet unidentified mechanism in patients dialyzed with high permeability membranes. the present study demonstrates that intact bacterial lipopolysaccharide (lps) molecules may cross cuprophan ... | 1990 | 2127434 |
| gentisate 1,2-dioxygenase from pseudomonas. purification, characterization, and comparison of the enzymes from pseudomonas testosteroni and pseudomonas acidovorans. | the 3-hydroxybenzoate inducible gentisate 1,2-dioxygenases have been purified to homogeneity from p. acidovorans and p. testosteroni, the two divergent species of the acidovorans group of pseudomonas. both enzymes exhibit a 40-fold higher specific activity than previous preparations and have an (alpha fe)4 quaternary structure (holoenzyme mr = 164,000 and 158,000, respectively). the enzymes have different amino terminal sequences, amino acid contents, and isoelectric points. each enzyme contains ... | 1990 | 2156846 |
| gentisate 1,2-dioxygenase from pseudomonas. substrate coordination to active site fe2+ and mechanism of turnover. | gentisate 1,2-dioxygenase catalyzes the oxygenolytic ring cleavage of gentisate (2,5-dihydroxybenzoate) between carbons 1 and 2 to form maleylpyruvate. the essential active site fe2+ of the enzyme binds no to yield an epr-active (s = 3/2) complex. hyperfine broadening from 17o (i = 5/2) is observed in the spectrum of the enzyme-nitrosyl complex prepared in 17o-enriched water, demonstrating that water is an iron ligand. association of gentisate with the enzyme-nitrosyl complex causes the broadeni ... | 1990 | 2266121 |
| protocatechuate 4,5-dioxygenase from pseudomonas testosteroni. | 1990 | 2280721 | |
| inhibition of microbial cholesterol oxidases by dimethylmorpholines. | cholesterol oxidase is a potentially important enzyme in steroid transformations, catalysing the conversion of 3-hydroxy-5-ene steroids to 3-keto-4-ene derivatives via a 3-keto-5-ene intermediate. morpholine derivatives, especially fenpropimorph and tridemorph, were found to block selectively the isomerisation activity of cholesterol oxidases isolated from nocardia erythropolis, streptomyces sp., pseudomonas testosteroni and schizophyllum commune. these enzymes differ strongly in physical charac ... | 1990 | 2308321 |
| cloning and expression of genes involved in 4-chlorobiphenyl transformation by pseudomonas testosteroni: homology to polychlorobiphenyl-degrading genes in other bacteria. | the genes of pseudomonas testosteroni strain b-356, specifying the transformation of 4-chlorobiphenyl (4-cb) into 4-chlorobenzoic acid (4-cba) were cloned into pseudomonas putida kt2440 using a broad-host-range cosmid, ppsa842. of 10,000 clones tested, four were able to transform 4-cb. gas chromatographic and mass spectrometric analysis of the catabolic products from two of the 4-cb-transforming clones carrying the hybrid plasmids, pda1 and pda2, demonstrated that pda1 carried a complete set of ... | 1990 | 2311936 |
| degradation of p-toluenesulphonic acid via sidechain oxidation, desulphonation and meta ring cleavage in pseudomonas (comamonas) testosteroni t-2. | pseudomonas (comamonas) testosteroni t-2 completely converted p-toluenesulphonic acid (ts) or p-sulphobenzoic acid (psb) to cell material, co2 and sulphate, with growth yields of about 5 g protein (mol c)-1. psb and sulphite were excreted as transient intermediates during growth in ts-salts medium. all reactions of a catabolic pathway involving sidechain oxidation and cleavage of the sulphonate moiety as sulphite were measurable in the soluble portion of cell extracts. degradation of ts and psb ... | 1989 | 2614395 |
| kinetic and ultraviolet spectroscopic studies of active-site mutants of delta 5-3-ketosteroid isomerase. | delta 5-3-ketosteroid isomerase (ec 5.3.3.1) of pseudomonas testosteroni promotes the highly efficient isomerization of delta 5-3-ketosteroids to delta 4-3-ketosteroids by means of a direct and stereospecific transfer of the 4 beta-proton to the 6 beta-position, via an enolic intermediate. an acidic residue responsible for the protonation of the 3-carbonyl function of the steroid and a basic group concerned with the proton transfer have been implicated in the catalytic mechanism. recent nmr stud ... | 1989 | 2706241 |
| [a sarcoma-static new species of pseudomonas, pseudomonas jinanensis sp. nov]. | a strain of gram negative bacteria was isolated from the surface soil of wuying hill at jinan, shandong province with gause's medium in 1973. it is a strain of antagonistic bacteria for hysterocervicoma, hepatoma and melanoma of mice screened from 2100 strains of bacteria. it is also antagonistic to staphylococcus aureus, bacillus subtilis and micrococcus. it is a gram negative bacterium with lophotrichous polar flagella. straight rods in shape or with a little slightly curved rods, 0.5-0.6 x 1- ... | 1989 | 2781786 |
| the laboratory diagnosis of peritonitis during continuous ambulatory peritoneal dialysis. | in 24 episodes of continuous ambulatory peritoneal dialysis associated peritonitis occurring in 21 patients, all dialysis fluids were cloudy and contained at least 100 cells mm-3, mostly polymorphs. gram-positive cocci were seen in centrifuge deposits from only nine of the fluids, but enrichment of 5 ml in liquoid and glucose broth yielded bacteria for all episodes, namely staphylococcus epidermidis (seven episodes), staph. aureus (six), streptococcus mitior (one), moraxella spp (four), acinetob ... | 1986 | 2871078 |
| positioning of a spin-labeled substrate analogue into the structure of delta 5-3-ketosteroid isomerase by combined kinetic, magnetic resonance, and x-ray diffraction methods. | we have shown by kinetic and magnetic resonance measurements that a spin-labeled substrate analogue, spiro[doxyl-2,3'-5' alpha-androstan]-17'beta-ol, binds at the substrate site of crystalline delta 5-3-ketosteroid isomerase (steroid delta-isomerase; ec 5.3.3.1) of pseudomonas testosteroni. the spin-labeled steroid is a linear competitive inhibitor with a ki value (25 +/- 5 microm) that is consistent with dissociation constants obtained by direct binding measurements based on changes in the elec ... | 1987 | 2888482 |
| lyophilized clostridium perfringens 3 alpha- and clostridium bifermentans 7 alpha-hydroxysteroid dehydrogenases: two new stable enzyme preparations for routine bile acid analysis. | preparations of 3 alpha-hydroxysteroid dehydrogenase (ec 1.1.1.50) from clostridium perfringens were successfully lyophilized into a stable powder form. purification of the enzyme was achieved using triazine dye affinity chromatography. c. perfringens 3 alpha-hydroxysteroid dehydrogenase was purified 24-fold using reactive red 120 (procion red) -cross-linked agarose (70% yield). quantitative measurement of bile acids with the purified enzymes, 3 alpha-hydroxysteroid dehydrogenase and 7 alpha-hyd ... | 1988 | 2901274 |
| isolation and characterization of a 50 kda testosterone-binding protein from pseudomonas testosteroni. | a testosterone-binding protein (mr = 50,500) has been isolated from the gram-negative bacterium pseudomonas testosteroni. the protein was partially purified by a combination of ion exchange chromatography and chromatofocusing. final purification was achieved by electroelution of the 50 kda protein from sds-polyacrylamide gels. following renaturation from a diluted solution of guanidine-hcl, specific binding of [3h]testosterone to the purified protein was observed. the native protein has a pi of ... | 1989 | 2913397 |
| 3 beta, 17 beta-hydroxysteroid dehydrogenase of pseudomonas testosteroni. kinetic evidence for the bifunctional activity at a common catalytic site. | 3 beta, 17 beta-hydroxysteroid dehydrogenase (3 beta 17 beta hsdh) is an nad-dependent dehydrogenase which has a double specificity for the 3- and 17-positions on the steroid skeleton. when dehydroepiandrosterone (dhea) is used as steroid substrate, and the assay coupled with ketosteroid-isomerase, the two reactions occur alternately and each reaction on the 3-position produces a chromophoric molecule. these two reactions can follow one another without dissociation of the coenzyme from the enzym ... | 1985 | 2991019 |
| [17o]water and nitric oxide binding by protocatechuate 4,5-dioxygenase and catechol 2,3-dioxygenase. evidence for binding of exogenous ligands to the active site fe2+ of extradiol dioxygenases. | pseudomonas testosteroni protocatechuate 4,5-dioxygenase and pseudomonas putida catechol 2,3-dioxygenase (metapyrocatechase) catalyze extradiol-type oxygenolytic cleavage of the aromatic ring of their substrates. the essential active site fe2+ of each enzyme binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complexes by 17o-enriched h2o shows that water is bound directly to the fe2+ in the nativ ... | 1985 | 2997190 |
| binding of 17o-labeled substrate and inhibitors to protocatechuate 4,5-dioxygenase-nitrosyl complex. evidence for direct substrate binding to the active site fe2+ of extradiol dioxygenases. | pseudomonas testosteroni protocatechuate 4,5-dioxygenase catalyzes extradiol-type oxygenolytic cleavage of the aromatic ring of its substrate. the essential active site fe2+ binds nitric oxide (no) to produce an epr active complex with an electronic spin of s = 3/2. hyperfine broadening of the epr resonances of the nitrosyl complex of the enzyme by protocatechuate (3,4-(oh)2-benzoate, pca) enriched specifically with 17o (i = 5/2) in either the 3 or the 4 hydroxyl group shows that both groups can ... | 1986 | 3003098 |
| inhibition of 3(17)beta-hydroxysteroid dehydrogenase from pseudomonas testosteroni by steroidal a ring fused pyrazoles. | several 2,3- and 3,4-steroidal fused pyrazoles have been investigated as potential inhibitors of nad(p)h-dependent steroid oxidoreductases. these compounds are proven to be potent, specific inhibitors for 3(17) beta-hydroxysteroid dehydrogenase from pseudomonas testosteroni with ki values of 6-100 nm. in contrast, the activities of 3 alpha,20 beta-hydroxysteroid dehydrogenase from streptomyces hydrogenans, steroid 5 alpha-reductase from rat prostate, and 3 alpha-hydroxysteroid dehydrogenase from ... | 1987 | 3040087 |
| new naphthalene-degrading marine pseudomonas strains. | over 100 strains that utilized naphthalene as the only carbon and energy source were isolated from samples of marine sediments taken from a heavily polluted area. the isolates were characterized taxonomically and physiologically. most of these strains belonged to the genus pseudomonas, and seven of them did not fit any previous taxonomic description. they differed from type strains in a few biochemical characteristics and in the utilization of aromatic compounds. none had catechol 1,2-dioxygenas ... | 1988 | 3202629 |
| nucleotide sequence of the gene for the delta 5-3-ketosteroid isomerase of pseudomonas testosteroni. | the structural gene for the delta 5-3-ketosteroid isomerase of pseudomonas testosteroni has been sequenced by the dideoxy method. the sequence obtained confirms the amino acid (aa) sequence of benson et al. [j. biol. chem. 246 (1971) 7514-7525] at all but 5 aa residues of the 125-aa polypeptide. amino acid residues 22, 24, 33, and 38, reported to be asparagines by benson et al., are found to be encoded by aspartic acid codons. amino acid residue 77, reported to be a glutamine by benson et al., i ... | 1988 | 3224818 |
| cloning of the gene for delta 5-3-ketosteroid isomerase from pseudomonas testosteroni. | we have cloned an approx. 5-kb fragment of pseudomonas testosteroni dna containing the structural gene of delta 5-3-ketosteroid isomerase into the ecori site of the lambda gt11 genome. escherichia coli infected with these recombinant phages produce a polypeptide which is recognized by antiserum raised against the purified isomerase. four of the recombinant lambda gt11 clones contain significant levels of isomerase activity and produce an immunopositive polypeptide of the same apparent mr as the ... | 1987 | 3428616 |
| isolation and sequencing of the gene encoding delta 5-3-ketosteroid isomerase of pseudomonas testosteroni: overexpression of the protein. | we describe the cloning, sequencing, and overexpression of the steroid isomerase (3-oxosteroid delta 5-delta 4-isomerase, ec 5.3.3.1) gene of pseudomonas testosteroni. a genomic library of p. testosteroni total dna constructed from partial ecori digests ligated to a lambda gtwes vector was probed with a 23-base oligonucleotide mixture [atgaac(t)acc(a,t)ccg(c,a)gag(a)cac(t)atgac] corresponding to the nh2-terminal sequence of steroid isomerase. subclones derived from a recombinant phage containing ... | 1987 | 3480517 |
| high affinity protein-binding and enzyme-inducing activity of methyltrienolone in pseudomonas testosteroni. | the synthetic androgen methyltrienolone (r 1881) was shown to increase steroid delta 1 dehydrogenase activity when added to cultures of pseudomonas testosteroni at concentrations of 10(-10)-10(-8)m. incubation with a soluble extract of p. testosteroni showed that (3h)-r 1881 was bound to a macromolecule with high affinity (kd 0.6 x 10(-9)m) and low capacity (number of binding sites 120 x 10(-15) mol/mg of protein). the (3h)-r 1881-macromolecule complex was partially destroyed following treatment ... | 1986 | 3490081 |
| quinohaemoprotein alcohol dehydrogenase apoenzyme from pseudomonas testosteroni. | cell-free extracts of pseudomonas testosteroni, grown on alcohols, contain quinoprotein alcohol dehydrogenase apoenzyme, as was demonstrated by the detection of dye-linked alcohol dehydrogenase activity after the addition of pqq (pyrroloquinoline quinone). the apoenzyme was purified to homogeneity, and the holoenzyme was characterized. primary alcohols (except methanol), secondary alcohols and aldehydes were substrates, and a broad range of dyes functioned as artificial electron acceptor. optima ... | 1986 | 3521592 |
| nad(p)+-independent aldehyde dehydrogenase from pseudomonas testosteroni. a novel type of molybdenum-containing hydroxylase. | aldehyde dehydrogenase from pseudomonas testosteroni was purified to homogeneity. the enzyme has a ph optimum of 8.2, uses a wide range of aldehydes as substrates and cationic dyes (wurster's blue, phenazine methosulphate and thionine), but not anionic dyes (ferricyanide and 2.6-dichloroindophenol), nad(p)+ or o2, as electron acceptors. haem c and pyrroloquinoline quinone appeared to be absent but the common cofactors of molybdenum hydroxylases were present. xanthine was not a substrate and allo ... | 1987 | 3609027 |
| affinity alkylation of 3-oxo-delta 5-steroid isomerase by steroidal 3 beta-oxiranes: identification of the modified amino acid by reduction with hydroxyborohydride. | the steroidal 3 beta-oxirane (3s)-spiro[5 alpha-androstane-3,2'-oxiran]-17 beta-ol (1 beta) is an active site directed irreversible inhibitor of the 3-oxo-delta 5-steroid isomerase from pseudomonas testosteroni. two steroid-bound peptides (tps1 and tps2) were isolated by high-performance liquid chromatography (hplc) from the trypsin digest of enzyme inactivated with 1 beta. the modified tryptic peptides (residues 14-45 of the enzyme) were further digested with chymotrypsin, each giving rise to a ... | 1987 | 3620446 |
| the amino acid sequence of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. | we have determined the primary structure of a delta 5-3-oxosteroid isomerase from pseudomonas putida biotype b. the enzyme is a dimeric protein of two identical subunits, each consisting of a polypeptide chain of 131 residues and a mr = 14,536. the intact s-carboxymethyl protein was sequenced from the nh2 terminus using standard automated edman degradation and automated edman degradation using fluorescamine treatment at known prolines to suppress background. the isomerase was fragmented using cn ... | 1986 | 3700400 |
| pseudomonas testosteroni infections: eighteen recent cases and a review of the literature. | pseudomonas testosteroni has been largely overlooked as a potential pathogen in humans. ten cases of infection due to p. testosteroni were identified at a single metropolitan hospital in texas during a three-year period. the organism was most often found in association with anatomic abnormalities of the gastrointestinal tract (six of 10 cases); perforation of the appendix was the commonest abnormality (five cases). the infections were more often polymicrobial (seven cases) than monomicrobial (th ... | 1987 | 3823716 |
| irreversible inhibition of delta 5-3-oxosteroid isomerase by 2-substituted progesterones. | 2 alpha-cyanoprogesterone (i) and 2-hydroxymethyleneprogesterone (ii) were synthesized and screened as irreversible active-site-directed inhibitors of the delta 5-3-oxosteroid isomerase (ec 5.3.3.1) from pseudomonas testosteroni. both compounds were found to inhibit the purified bacterial enzyme in a time-dependent manner. in either case the inactivated enzyme could be dialysed without return of activity, indicating that a stable covalent bond had formed between the inhibitor and the enzyme. ina ... | 1985 | 3838891 |
| mitochondrial origins. | the 16s ribosomal rna sequences from agrobacterium tumefaciens and pseudomonas testosteroni have been determined to further delimit the origin of the endosymbiont that gave rise to the mitochondrion. these two prokaryotes represent the alpha and beta subdivisions, respectively, of the so-called purple bacteria. the endosymbiont that gave rise to the mitochondrion belonged to the alpha subdivision, a group that also contains the rhizobacteria, the agrobacteria, and the rickettsias--all prokaryote ... | 1985 | 3892535 |
| mechanism of inactivation of 3-oxosteroid delta 5-isomerase by 17 beta-oxiranes. | the affinity label (17s)-spiro[estra-1,3,5(10),6,8-pentaene-17,2'-oxiran]-3-ol (5 beta) inactivates 3-oxosteroid delta 5-isomerase from pseudomonas testosteroni by formation of a covalent bond between asp-38 of the enzyme and the steroid. high-performance liquid chromatography (hplc) analysis of tryptic digests of inactivated enzyme shows that two isomeric steroid-containing peptides are formed in a ratio of 9:1 at ph 7 (tps1 and tps2). hydrolysis of each of these peptides produces a different s ... | 1985 | 4027215 |